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Open Access Research article

Effect of cAMP derivates on assembly and maintenance of tight junctions in human umbilical vein endothelial cells

Michaela Beese, Kristin Wyss, Marion Haubitz and Torsten Kirsch*

Author Affiliations

Division of Nephrology, Department of Internal Medicine, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany

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BMC Cell Biology 2010, 11:68  doi:10.1186/1471-2121-11-68

Published: 7 September 2010

Additional files

Additional file 1:

Immunostainings of VE-cadherin. HUVEC were stimulated with cAMP or different cAMP derivates for 24 to 72 h and stained for VE-cadherin. The nucleus was counterstained with DAPI. Shown are representative confocal images of at least three independent experiments with HUVEC derived from different donors. Scale bar represents 25 μm.

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Additional file 2:

Immunostainings of tight junction marker after 48 h stimulation with cAMP derivates. Shown are representative micrographs of immunostainings for ZO-1, OCLN, Jam-A and CLDN5 in HUVEC that were stimulated with cAMP or its derivates for 48 h. The nucleus was counterstained with DAPI. Scale bar represents 25 μm.

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Additional file 3:

mRNA expression data of junction-associated proteins. HUVEC were stimulated for 24 to 72 h with cAMP or its derivates. RNA was extracted, reverse transcribed and expression of VE-cadherin, ZO-1 and Jam-A was determined by real-time qPCR. Expression data were normalized to relative transcript amounts of HPRT-1 (n = 3).

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Additional file 4:

cAMP-induced expression of CLDN5 and OCLN is mediated by PKA. (A) HUVEC were pretreated with H-89 for 30 minutes and stimulated with cAMP or its derivates for up to 72 h. mRNA expression of OCLN and CLDN5 were determined by real-time qPCR and normalized to expression of HPRT-1 (n = 3). (B) Shown are representative western blot images of HUVEC pretreated with H-89 for 30 minutes and stimulated with cAMP derivates for up to 72 h. Protein amounts of tubulin served as loading control (n = 2).

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